ABSTRACT
Objective:To optimize the major parameters of bioinformatics analysis conditions in process of PMF(Peptide Mass Fingerprinting) identify target proteins. Methods:In this process of experiment design and analysis,we made the Bovine Carbonic anhydrase-2 and BSA from 2-DE gel,after proteolysis with porcine trypsin and gathered the peptides,subsequently identified with MALDI-TOF MS and got the peptides mass data. Then,we selected database of Swissprot and MS-Fit search engine,the standard Carbonic anhydrase-2 protein used as model to set up protein search standard conditions. Results:Our study showed that the cysteine was modified by carbamidomethylation more convenient in protein search,at the same time,the better mode of mass tolerance was percentage and 0.1% was suitable tolerance in our search. Then we used BSA mass data to check the accuracy of MS search standard conditions. Conclusions:Results show that the optimized parameters are reliable.
ABSTRACT
Proteomic technology has been widely used in the research of differently-expressed proteins under different physiology and pathologic conditions.In the aspect of the human spermatozoa proteomics,many reports have demonstrated the differently expressed proteins and the roles of sperm under different physiology and pathologic state from proteome level,and these results provided reliable theoretic base for well understanding of sperm molecule mechanism in physiology and pathologic conditions.